高效液相色谱法灵香草中槲皮素含量

Determination of Quercetin in Extract from Lysimachia foenum-graecum Hance by HPLC

为了测定灵香草中槲皮素含量,以2 mol/L的盐酸甲醇溶液为提取溶剂,采用Wondasil C18柱(4.6 mm×250 mm,5μm)为分离柱,筛选流动相甲苯、乙酸乙酯和甲酸的不同配比,探索HPLC法测定灵香草甲醇提取物中槲皮素含量的最佳条件.结果显示,当甲苯、乙酸乙酯和甲酸的配比为10:8:1,流速为1.5 mL/min时槲皮素分离效果较好;紫外检测波长为375 nm时,槲皮素保留时间为19 min;以槲皮素保留时间和峰面积进行定性与定量分析,峰面积与含量绝对相关系数是0.9 998,相对标准偏差(RSD)变化范围为1.83%~2.49%,加样回收率平均为96.44%,表明采用HPLC法检测灵香草甲醇提取物中槲皮素含量灵敏度高、可重复性好.本研究可为灵香草药材成分的开发利用提供较好的参考.

High-performance liquid chromatography(HPLC)was developed for the separation of quercetin from the methanol extracts fromLysimachia foenum-graecum Hance.The quercetin was extracted using the solvent of 2 mol/L methanol-HCl.Chromatographic separation was performed in Wondasil C18(4.6 mm×250 mm,5μm)with an eluent consisting of methylbenzene,ethyl acetate,formic acid,the volume ratio of the three kinds of reagent was 10∶8∶1(by volume),at a flow rate of 1.5 mL/min,the eluate was monitored at 375 nm.The result showed that separation and identification of quercetin were completed in 30 min.The linear ranges of quercetin were 0.0250~0.0750mg/mL(R^2=0.9998).This method was precise and accurate(RSD 1.83%~2.49%),the average recoveries percent was 96.44%.HPLC is simple,accurate for check of quercetin of Lysimachia foenum-graecum Hance.This study provides a good reference for the development and utilization of Lysimachia foenum-graecum Hance.